CST, 14989S, PTMScan® Phospho-Ser-Pro-Pro Motif [pSPP] Kit

PTMScan for studying in the research area. Product Usage Information Cells are lysed in a urea-containing buffer, cellular proteins are digested by proteases, and the resulting peptides are purified by reversed-phase solid-phase extraction. Peptides are then subjected to immunoaffinity purification using a PTMScan ® Motif Antibody conjugated to protein A agarose beads. Unbound peptides are removed through washing, and the captured PTM-containing peptides are eluted with dilute acid. Reversed-phase purification is performed on microtips to desalt and separate peptides from antibody prior to concentrating the enriched peptides for LC-MS/MS analysis. CST recommends the use of PTMScan ® IAP Buffer #9993 included in the kit. Storage Antibody beads supplied in IAP buffer containing 50% glycerol. Store at -20ºC. Do not aliquot the antibody. Protocol Available protocols: PTMScan Background The MAPK and CDK families of serine/threonine protein kinases play important roles in cell signaling and cell cycle control. These kinases phosphorylate threonine or serine followed by a proline residue (1-6). To facilitate the study and discovery of new MAPK and CDK substrates, Cell Signaling Technology has developed antibodies that bind to phospho-threonine or phospho-serine followed by proline. The S/TPP motif matches the consensus sequence for targeted phosphorylation by Cdc2 (7,8), ERK1, ERK2 (9), SAPK/JNK, CDK5, and GSK3 (10). Alternate Names SPP